췌관내 유두상 점액종에서의 마이크로 RNA 발현 양상

박윤경 ( Yun Gyoung Park ),이광혁 ( Kwang Hyuck Lee ),이종균 ( Jong Kyun Lee ),이규택 ( Kyu Taek Lee ),최동욱 ( Dong Wook Choi ),최성호 ( Seong Ho Choi ),허진석 ( Jin Seok Heo ),장기택 ( Kee Taek Jang ),이은미 ( Eun Mi Lee ),김정 대한소화기학회 2011 대한소화기학회지 Vol.58 No.4

Background/Aims: Intraductal papillary mucinous neoplasms (IPMN) are precursor lesions of fatal pancreatic cancer. Physiological function of microRNA is to regulate the stability and translation of mRNA. The aberrant microRNA expression is commonly observed in many cancers. The aim of this study was to analyze the expression pattern of microRNA in IPMN and evaluate the role of the microRNA. Methods: Using two paraffin-embedded IPMN tissues, microRNA expression of normal tissue, IPMN adenoma and carcinoma were compared by cDNA-mediated annealing, selection, extension and ligation microarray assay. Using real time PCR, expression levels of aberrantly up-regulated microRNAs were assessed in another 20 IPMNs, four pancreatic cancer cell lines (Panc1, MiaPaCa-2, XPA-3, BxPC-3) and immortalized pancreatic ductal cell line (HPNE). Effect of suppressing highly over-expressed two microRNAs in pancreatic cancer cell lines with anti-microRNA inhibitors were evaluated using CCK-8 assay. Results: Among aberrantly expressed 122 microRNAs in IPMN, miR-552, miR-25*, miR-183, miR-1300, miR-196a, miR-182*, and miR-30c-1* were consistently increased more than 3-fold. On average, miR-196a and miR-183 increased 10,824 folds and 26,519 folds in four pancreatic cancer cell lines compared with HPNE. These two microRNAs were also over-expressed in 20 IPMNs compared with HPNE. After applying anti-miRNA inhibitors, cell survival of four pancreatic cancer cell lines decreased by 24.5% with anti-miR-196a and by 14.2% with anti-miR-183 on average. Conclusions: Aberrant expression of 122 microRNAs was observed in IPMN. Two microRNAs, miR-196a and miR-183-increased in IPMN and pancreatic cancer cell lines compared with immortalized dancreatic ductal cell line. The inhibitions of these microRNAs repressed cell proliferation of pancreatic cancer cell lines. (Korean J Gastroenterol 2011;58:190-200)

MicroRNA profiling of tacrolimus-stimulated Jurkat human T lympocytes

Ho Kyun Lee,Sang Young Chung,Soo Jin Na Choi 대한외과학회 2013 Annals of Surgical Treatment and Research(ASRT) Vol.85 No.4

Purpose: This study investigated the Jurkat T cell line expresses cytotoxicity when treated with different concentrations of FK506, and analyzed the expression pattern of microRNA when stimulated by FK506 using the microRNAs microarray, as well as the expression pattern of a gene that is related to the differentiation, activation and proliferation of T cells after being affected by the change of microRNAs. Methods: To investigate the effects of FK506 on microRNA expression, we purified total RNA of Jurkat cells treated with 20 μM FK506 for 72 hours and used to analyze microRNA profiling by using Agilent’s chip. Results: These results demonstrated that treatment with FK506 markedly induced the down-regulation of 20 microRNAs as well as the up-regulation of 20 microRNAs in a time-dependent manner. The genes that down-regulated by FK506 include let-7a*, miR-20a*, and miR-487a. Otherwise miR-202, miR-485-5p, and miR-518c* are gradually up-regulated in expression. Sanger Institute and DAVIDs bioinformatics indicated that microRNAs regulated the several transcriptomes including nuclear factor of activated T cell-related, T cell receptor/interleukin-2 signaling, and Ca<SUP>2+</SUP>-calmodulin-dependent phosphatase calcineurin pathways. Conclusion: As a result of treating FK506 to a Jurkat cell line and running the microRNA microarray, it was found that FK506 not only took part in the suppression of T cell proliferation/activation by inhibiting calcineurin in Jurkat apoptosis, but also affected the microRNAs that are involved in the regulation of various signal transduction pathways.

BRCA1 and MicroRNAs: Emerging Networks and Potential Therapeutic Targets

Suhwan Chang,Shyam K. Sharan 한국분자세포생물학회 2012 Molecules and cells Vol.34 No.5

BRCA1 is a well-known tumor suppressor implicated in familial breast and ovarian cancer. Since its cloning in 1994, numerous studies have established BRCA1’s role in diverse cellular and biochemical processes, such as DNA damage repair, cell cycle control, and transcriptional regulation as well as ubiquitination. In addition, a number of recent studies have functionally linked this tumor suppressor to another important cellular regulator, microRNAs, which are short (19-22 nt) RNAs that were discovered in the nematode in 1993. Soon their presence and function were validated in mammals, and since then, the role of microRNAs has been actively investigated in almost all biological processes, including cancer. In this review, we will describe recent progress in the understanding of the BRCA1 function through microRNAs and the role of microRNAs in regulating BRCA1, with emphasis on the implication of these processes on the development and progression of cancer. We will also discuss the therapeutic potential of microRNA mimics or inhibitors of microRNAs to affect BRCA1 function.

Urinary and Blood MicroRNA-126 and -770 are Potential Noninvasive Biomarker Candidates for Diabetic Nephropathy: a Meta-Analysis

Park, Sungjin,Moon, SeongRyeol,Lee, Kiyoung,Park, Ie Byung,Lee, Dae Ho,Nam, Seungyoon S. Karger AG 2018 CELLULAR PHYSIOLOGY AND BIOCHEMISTRY Vol.46 No.4

<P><B><I>Background/Aims:</I></B> Diabetic nephropathy (DN), a major diabetic microvascular complication, has a long and growing list of biomarkers, including microRNA biomarkers, which have not been consistent across preclinical and clinical studies. This meta-analysis aims to identify significant blood- and urine-incident microRNAs as diagnostic/prognostic biomarker candidates for DN. <B><I>Methods:</I></B> PubMed, Web of Science, and Cochrane Library were searched from their earliest records through 12th Dec 2016. Relevant publications for the meta-analysis included (1) human participants; (2) microRNAs in blood and urine; (3) DN studies; and (4) English language. Four reviewers, including two physicians, independently and blindly extracted published data regarding microRNA profiles in blood and/or urine from subjects with diabetic nephropathy. A random-effect model was used to pool the data. Statistical associations between diabetic nephropathy and urinary or blood microRNA expression levels were assessed. <B><I>Results:</I></B> Fourteen out of 327 studies (n=2,747 patients) were selected. Blood or urinary microRNA expression data of diabetic nephropathy were pooled for this analysis. The hsa-miR-126 family was significantly (OR: 0.57; 95% CI: 0.44-0.74; p-value < 0.0001) downregulated in blood from patients with diabetic kidney disease, while its urinary level was upregulated (OR: 2931.12; 95% CI: 9.96-862623.21; p-value = 0.0059). The hsa-miR-770 family microRNA were significantly (OR: 10.24; 95% CI: 2.37-44.25; p-value = 0.0018) upregulated in both blood and urine from patients with diabetic nephropathy. <B><I>Conclusions:</I></B> Our meta-analysis suggests that hsa-miR-126 and hsa-miR-770 family microRNA may have important diagnostic and pathogenetic implications for DN, which warrants further systematic clinical studies.</P>

Cell-Free microRNA-214 From Urine as a Biomarker for Non–Muscle-Invasive Bladder Cancer

김성민,강호원,김원태,김용준,윤석중,이상철,김원재 대한비뇨의학회 2013 Investigative and Clinical Urology Vol.54 No.11

Purpose: MicroRNAs are small noncoding RNAs and microRNA-214 (miR-214) has been associated with the inhibition of cancer cell growth, migration, and invasion. The aim of this study was to investigate whether cell-free miR-214 isolated from urine could be used as a biomarker for non–muscle-invasive bladder cancer (NMIBC). Materials and Methods: A total of 138 patients with primary NMIBC and 144 healthy normal controls were enrolled in this study. By use of quantitative polymerase chain reaction (PCR), the urinary levels of cell-free miR-214 were measured and the clinicopathological parameters of patients with NMIBC were compared with those of the controls. Results: The urinary levels of cell-free miR-214 were significantly higher in the NMIBC patients than in the controls (20.08±3.21 vs. 18.96±2.68, p=0.002). However, the urinary levels of cell-free miR-214 were neither graded nor staged for the NMIBC patients (p>0.05, each). When we compared the urinary levels of cell-free miR-214 according to clinical outcomes, patients with recurrence had lower levels of miR-214 than did those with no recurrence (19.24±2.67 vs. 20.41±3.41, p=0.023). By contrast, there were no significant differences in the urinary level of cell-free miR-214 between the NMIBC patients showing progression and those showing no progression (p=0.919). Multivariate Cox regression analysis revealed that urinary levels of cell-free miR-214 were an independent predictor of NMIBC recurrence (hazard ratio, 2.011; 95% confidence interval, 1.027 to 3.937; p=0.041). Conclusions: Urinary levels of cell-free miR-214 could be an independent prognostic parameter for NMIBC recurrence. Thus, urinary cell-free microRNA-214 might be a useful prognostic marker for NMI BC. Purpose: MicroRNAs are small noncoding RNAs and microRNA-214 (miR-214) has been associated with the inhibition of cancer cell growth, migration, and invasion. The aim of this study was to investigate whether cell-free miR-214 isolated from urine could be used as a biomarker for non–muscle-invasive bladder cancer (NMIBC). Materials and Methods: A total of 138 patients with primary NMIBC and 144 healthy normal controls were enrolled in this study. By use of quantitative polymerase chain reaction (PCR), the urinary levels of cell-free miR-214 were measured and the clinicopathological parameters of patients with NMIBC were compared with those of the controls. Results: The urinary levels of cell-free miR-214 were significantly higher in the NMIBC patients than in the controls (20.08±3.21 vs. 18.96±2.68, p=0.002). However, the urinary levels of cell-free miR-214 were neither graded nor staged for the NMIBC patients (p>0.05, each). When we compared the urinary levels of cell-free miR-214 according to clinical outcomes, patients with recurrence had lower levels of miR-214 than did those with no recurrence (19.24±2.67 vs. 20.41±3.41, p=0.023). By contrast, there were no significant differences in the urinary level of cell-free miR-214 between the NMIBC patients showing progression and those showing no progression (p=0.919). Multivariate Cox regression analysis revealed that urinary levels of cell-free miR-214 were an independent predictor of NMIBC recurrence (hazard ratio, 2.011; 95% confidence interval, 1.027 to 3.937; p=0.041). Conclusions: Urinary levels of cell-free miR-214 could be an independent prognostic parameter for NMIBC recurrence. Thus, urinary cell-free microRNA-214 might be a useful prognostic marker for NMI BC.

Investigation of Dysregulation of Several MicroRNAs in Peripheral Blood of Schizophrenia Patients

Mehmet Akif Camkurt,Fatih Karababa,Mehmet Emin Erdal,Hüseyin Bayazıt,Sultan Basmacı Kandemir,Mustafa Ertan Ay,Hasan Kandemir,Özlem İzci Ay,Erdinç Çiçek,Salih Selek,Bahar Taşdelen 대한정신약물학회 2016 CLINICAL PSYCHOPHARMACOLOGY AND NEUROSCIENCE Vol.14 No.3

Objective: The prevalence of schizophrenia is 1%, and it is a debilitating disorder that often results in a shortened lifespan. Peripheral blood samples are good candidates to investigate because they can be easily drawn, and they are widely studied in psychiatric disorders. MicroRNAs are small non-coding RNA transcripts. They regulate the expression of genes by binding to the 3’-untranslated region (UTR) of mRNAs and pointing them to degrade. In this study, we aimed to investigate the expression of miR-9-5p, miR-29a-3p, miR-106-5p, miR-106b-5p, miR-107, miR-125a-3p, and miR-125b-3p in schizophrenia patients and healthy controls. Methods: We collected blood samples from 16 patients with schizophrenia and 16 healthy controls. MicroRNAs were measured with reverse transcriptase polymerase chain reaction. Results: Schizophrenia patients showed statistically significant upregulation of five microRNAs: miR9-5p (p=0.002), miR29a-3p (p<0.001), miR106b-5p (p=0.002), miR125a-3p (p<0.001), and miR125b-3p (p=0.018). Conclusion: Our results increased the value of the miR106 and miR29 families as potentially and consistently dysregulated in psychiatric disorders. Our results should be considered preliminary, and they need confirmation in future studies with larger sample sizes.

Expression of MicroRNA-221 in Korean Patients with Multiple Myeloma

최우순,Choi, Woo-Soon Korean Society for Clinical Laboratory Science 2018 대한임상검사과학회지(KJCLS) Vol.50 No.2

다발성 골수종(MM)은 혈액 종양의 주요 사망 원인이다. 최근에 다발성 골수종 진단에 microRNA를 이용한 실험이 보고되고 있다. 이에 우리는 다발성 골수종 진단 마커로 miR-221을 이용 할 수 있는지 확인하고자 하였다. 본 연구는 다른 혈액학적 질환이 없는 다발성 골수종 환자 20명을 대상으로 하였다. MicroRNA 추출은 다발성 골수종 환자의 파라핀 포매 조직을 이용하였다. 우리는 다발성 골수종의 microRNA 표적 유전자로 miR-15a, miR-16, miR-21, miR-181a 그리고 miR-221을 선택하였다. 유의성 검정은 fold change 값을 기준으로 1.5이상 또는 -1.5 미만의 결과를 유의성이 있는 경우로 하였다. Fold change 값은 인간 유전자 SNORD43에 의해 표준화된 데이터를 기준으로 하였다. Fold change 값이 1.5 이상은 "overexpression", -1.5 미만의 값은 "underexpression"으로 정의되었다. miR-221의 65.0% (13/20)에서 "overexpression"으로 유의성이 있음을 확인하였고, 다발성 골수종 환자에서 형질세포가 30% 이상인 그룹과 이하의 그룹은 유의성을 보이지 않았다. MiR-221은 서구인과 같은 결과를 얻었으며, 다발성 골수종 환자에서 miR-221이 다발성 골수종 환자 진단에 매우 중요한 지표가 될 수 있을 것으로 생각된다. 결론적으로 miR-221은 한국인의 다발성 골수종 진단 또는 예후 지표로 활용할 수 있음을 확인하였다. Multiple myeloma (MM) is the leading cause of death among hematologic neoplasms. Recently, microRNA has been reported to be useful in the diagnosis of multiple myeloma. This study examined whether miR-221 could be used as a diagnostic marker for multiple myeloma. The study was performed on 20 patients with multiple myeloma without any other hematological diseases. MicroRNA extraction was performed using formalin-fixed paraffin-embedded (FFPE) tissues obtained from the bone marrow of patients with multiple myeloma. miR-15a, miR-16, miR-21, miR-181a, and miR-221 were selected as the microRNA target genes for multiple myeloma. The significance of microRNA was based on a fold change of <1.5. To quantify the fold changes, data normalized to the human gene, SNORD43, were used as the values of the patient group. Fold change values greater than 1.5 were defined as "overexpression", whereas values less than -1.5 were defined as "underexpression". Of note, 65.0% (13/20) of samples showed significant "overexpression" in the levels of miR-221 expression and plasma cells with a group of more and less than 30% in MM patients did not show any significance of plasma cell (P<0.05). The results of other studies showing a correlation between the expression of miR-221 and MM in Caucasians were confirmed. These results suggest that miR-221 may be a useful indicator for diagnosing patients with MM. In conclusion, miR-221 is useful in the diagnosis and determining the prognosis of multiple myeloma in Koreans.

Inhibition of miR-146a-5p and miR-8114 in Insulin-Secreting Cells Contributes to the Protection of Melatonin against Stearic Acid-Induced Cellular Senescence by Targeting Mafa

Shenghan Su,Qingrui Zhao,Lingfeng Dan,Yuqing Lin,Xuebei Li,Yunjin Zhang,Chunxiao Yang,Yimeng Dong,Xiaohan Li,Romano Regazzi,Changhao Sun,Xia Chu,Huimin Lu 대한내분비학회 2022 Endocrinology and metabolism Vol.37 No.6

Background: Chronic exposure to elevated levels of saturated fatty acids results in pancreatic β-cell senescence. However, targets and effective agents for preventing stearic acid-induced β-cell senescence are still lacking. Although melatonin administration can protect β-cells against lipotoxicity through anti-senescence processes, the precise underlying mechanisms still need to be explored. Therefore, we investigated the anti-senescence effect of melatonin on stearic acid-treated mouse β-cells and elucidated the possible role of microRNAs in this process. Methods: β-Cell senescence was identified by measuring the expression of senescence-related genes and senescence-associated β-galactosidase staining. Gain- and loss-of-function approaches were used to investigate the involvement of microRNAs in stearic acid-evoked β-cell senescence and dysfunction. Bioinformatics analyses and luciferase reporter activity assays were applied to predict the direct targets of microRNAs. Results: Long-term exposure to a high concentration of stearic acid-induced senescence and upregulated miR-146a-5p and miR-8114 expression in both mouse islets and β-TC6 cell lines. Melatonin effectively suppressed this process and reduced the levels of these two miRNAs. A remarkable reversibility of stearic acid-induced β-cell senescence and dysfunction was observed after silencing miR-146a-5p and miR-8114. Moreover, V-maf musculoaponeurotic fibrosarcoma oncogene homolog A (Mafa) was verified as a direct target of miR-146a-5p and miR-8114. Melatonin also significantly ameliorated senescence and dysfunction in miR-146a-5pand miR-8114-transfected β-cells. Conclusion: These data demonstrate that melatonin protects against stearic acid-induced β-cell senescence by inhibiting miR-146a-5p and miR-8114 and upregulating Mafa expression. This not only provides novel targets for preventing stearic acid-induced β-cell dysfunction, but also points to melatonin as a promising drug to combat type 2 diabetes progression.

Next generation sequencing analysis of exosomal microRNAs: Fusobacterium nucleatum regulates expression profiling of exosomal microRNAs in human colorectal cancer cells

Mi Ra Yu,Hye Jung Kim,Ji Wan Kang,Yun Hak Kim,Hae Ryoun Park 대한구강생물학회 2020 International Journal of Oral Biology Vol.45 No.3

Colon cancer is one of the most common malignant tumors, but there are still a few validated biomarkers of colon cancer. Exosome-mediated microRNAs (miRNAs) have been recognized as potential biomarkers in cancers, and miRNAs can regulate a variety of genes. Recently, Fusobacterium nucleatum was discovered in the tissues of human colon cancer patients. Its role in colon cancer was highlighted. F. nucleatum may contribute to the progression of colon cancer through the mechanism of exosome-mediated miRNAs transfer. However, the exosomal miRNAs regulation mechanism by F. nucleatum in colon cancer is not well known. Thus, we performed next-generation sequencing to investigate the overall pattern of exosomal miRNAs expression in the colon cancer cell culture supernatant. We have confirmed the alterations of various exosomal miRNAs. In addition, to investigate the function of exosomal miRNAs, a Kyoto Encyclopedia of Genes and Genomes analysis was performed on the target genes of changed miRNAs. Potential target genes were associated with a variety of signaling pathways, and one of these pathways was related to colorectal cancer. These findings suggested that F. nucleatum can alter exosomal miRNAs released from colorectal cancer cells. Furthermore, exosomal miRNAs altered by F. nucleatum could be potential biomarkers for the diagnosis and therapy of colon cancer.

췌장암에 대한 예후 생물표지자로서의 MicroRNA-200c

백우현 ( Woo Hyun Paik ),송병준 ( Byeong Jun Song ),김형우 ( Hyoung Woo Kim ),김혜리 ( Hye Ree Kim ),황진혁 ( Jin Hyeok Hwang ) 대한소화기학회 2015 대한소화기학회지 Vol.66 No.4

Background/Aims: MicroRNA (miRNA) regulates messenger RNA stability and translation. In cancer biology, miRNA affects the growth and metastasis of cancer cells by controlling epithelial-mesenchymal transition (EMT). MiR-200 family (200a/200b/ 200c/141) and miR-205 are associated with the regulation of EMT. We investigated the prognostic role of EMT-related miRNAs in pancreatic cancer. Methods: We analyzed miR-200 family and miR-205 expression in tissue samples of 84 patients who underwent radical resection for pancreatic cancer. Results: Patients were followed from the date of diagnosis until death or censoring. The mean overall survival was 25.0±2.0 months (2-140 months). The R0 resection rate was obtained in 84.5% (n=71) of patients. The relative expressions of miR-200a/200b/200c/141 and miR-205 were 266.9±57.3/18.5±2.2/0.7±0.1/27.2±6.6 folds and 0.1±0.1 compared with human pancreatic ductal epithelial cells, respectively. Overall survival was longer in the low miR-200c expression group than in the high expression group (35 vs. 19 months, p=0.013). Multivariate analysis confirmed that patients with low miR-200c expression survived longer than the high expression group (hazard ratio, 1.771, 95% CI, 1.081-2.900, p=0.023). There was a trend toward longer disease-free survival in low miR-200c group without statistical significance (p=0.061). Conclusions: The expression of miR-200c may be an important prognosis factor in pancreatic cancer, and it could be a novel therapeutic target of pancreatic cancer. (Korean J Gastroenterol 2015,66:215-220)